• Journal of Environmental Health Sciences
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• v.27 no.3
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• pp.63-70
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• 2001

To identify and evaluate the benzidine-DNA adducts in liver and bladder, we exposed the 80 ppm benzidine to 40 sprague-dawley rats by drinking water for 4 weeks(6.2 mg/kg body wt./day). Only one benzidine-DNA adduct was found at the same site of thin layer chromatogram of $^{32}$ P-postlabeling method in the liver and bladder of exposed rats. So we know the DNA adduct formed at liver and bladder were similar to each other, which was N-(deoxyguanosin-8-yl)-N'-acetylbenzidine. Relative adduct labeling(RAL) of DNA adduct was similar to each other for 1 and 2 weeks, but that in liver was significantly higher than in bladder for 3 and 4 weeks. RAL$\times$10$^{9}$ of DNA adduct were 84.45$\pm$11.31 and 152.8$\pm$5.53 in liver, and were 24.76$\pm$7.06 and 38.00$\pm$10.57 in bladder for 3 and 4 weeks, respectively. Regression equation between liver and bladder was Y=-3.801+2.507 X(r=0.6036, p<0.01). In conclusion, benzidine-DNA adduct formed in liver was significantly higher than that in bladder, with the similar compound structure in sparague-dewley rates treated benzidine in drinking water.

• Journal of Environmental Health Sciences
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• v.28 no.4
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• pp.6-11
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• 2002

3.3'-Dichlorobenzidine(DCB) has be shown carcinogenic in several animals, and the development of non-invasive biomonitoring method in workers exposed with it is a very important subject. DNA adduct is a good biomarker for biomonitoring about carcinogens exposure, and lymphocytes is a good non-invasive samples. So we studied to analyze metabolites in blood lymphocytes of female Sprague-Dawley rats exposed orally with DCB(20, 30, and 40 mg/kg wt.) for 3 weeks. For analysis of them, we isolated DNA adducts from blood lymphocytes by using the enzymes method in /sup 32/P-postlabeling, and measured them by using gas chromatography/mass spectrometry-selected ion monitoring(GC/MS-SIM). 4-aminobiphenyl and phenanthrene-d/sub 10/ were added as internal standard for blank sample. Standard metabolites of DCB were synthesized with using pyridine and acetic acid which were promoter and controller in acetylation of DCB. And they were used for calibration curve. Our results showed two kinds of metabolites in DNA adducts of blood lymphocytes. They were N-acetyl 3,3'-dichlorobenzidine(acDCB) and N,N'-diacetyl 3,3'-dichiorobenzidine(di-acDCB ). They were combined with DNA at the same time as an acetyl of it was removed. So we measured DCB and acDCB for two kinds of metabolites in DNA adducts of blood lymphocytes. Our results showed the levels of DCB were 1.46∼2.26 times more than that of acDCB. And also the levels of metabolites in 20, 30 and 40 mg/kg wt. were gradually increased with going days from 1st to 3rd week. They are 1.66, 1.38 and 0.90 times in total metabolites, 1.76, 1.49 and 1.02 times in DCB, and 1.51, 1.22 and 1.28 times in acDCB. In conclusion, the results of this study showed DCB exposed to rats formed DNA adduct in blood lymphocytes after acetylated to N-acetyl 3.3'-dichloro benzidine(acDCB) and N,N'-diacetyl 3,3'-dichlorobenzidine(di-acDCB), and they could be analyzed by us ing gas chromatography/mass spectrometry-selected ion monitoring(GC/MS-SIM).

• Toxicological Research
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• v.6 no.1
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• pp.63-73
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• 1990

Human exposure to environmental carcinogens can be detected by a number of methods including immunoassay, $^{32}P-postlabeling$ assay, and fluorescence technique. These assays have been applied to measure biological markers of carcinogen-adducts formed with macromolecules such as DNA, RNA and protein. In an attempt to investigate causal relationships between carcinogen exposure and tumor formation, specific carcinogen-adducts have been quantitated from human tissues and body fluids of cancer patients, occupational workers heavily exposed to certain carcinogens, smokers and controls. Carcinogens studied for biological human monitoring include benzo(a)pyrene, aflatoxin B1, UV light, ethylene oxide, 8-methoxypsoralen, 4-aminobiphenyl, vinyl choride, N-nitrosamine, cisplatin and other chemotherapeutic agents. Relevance of human monitoring for cancer research, progress in this field, methods to detect carcinogen-adducts are reviewed here. It is hoped that these approaches will be used for the risk assessment of carcinogen exposure, cancer etiology study and cancer prevention in humans.

• Toxicological Research
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• v.6 no.1
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• pp.61-61
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• 1990

Human exposure to environmental carcinogens can be detected by a number of methods including immunoassay, $^{32}P$-postlabeling assay, and fluorescence technique. These assays have been applied to measure biological markers of carcinogen-adducts formed with macromolecules such as DNA, RNA and protein. In an attempt to investigate causal relation ships between carcinogen exposure and tumor formation, specific carcinogen-adducts have been quantitated from human tissues and body fluids of cancer patients, occupational workers heavily exposed to certain carcinogens, smokers and controls. Carcinogens studied for biological human monitoring include benzo(a)pyrene, aflatoxin B1, UV light, ethylene oxide, 8-methoxypsoralen, 4-aminobiphenyl, vinyl chloride, N-nitrosamine, cisplatin and other chemotherapeutic agents. Relevance of human monitoring for cancer research, progress in this field, methods to detect carcinogen-adducts are reviewed here. It is hoped that these approaches will be used for the risk assessment of carcinogen exposure, cancer etiology study and cancer prevention in humans.

• Environmental Mutagens and Carcinogens
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• v.17 no.2
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• pp.97-108
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• 1997

The drugs or xenobiotics introduced to the body, are detoxified through the process of biotransformation in the body. In this process, most of the insoluble compounds become more polar, soluble and easily excretable. But, parts of introduced materials are metabolized to highly reactive electrophilic carcinogens through activation pathways. These metabolites are toxic and can react with DNA, RNA and proteins which are nucleophilic compounds. The objective of this study is to illustrate the aleactivation pathways of two highly reactive epoxide compounds, vinyl carbamate epoxide (VCO) and 2'-(4-nitrophenoxy)oxirane (NPO). They are the ultimate electrophilic carcinogens of ethyl carbamate(urethane) and 4-nitrophenyl vinyl ether, respectively. In this research, we studied the inhibition of the mutagenic activities of VCO or NPO by nuchieophiles [glutahione(GSH) and N-acetylcysteine(NAC)], detoxifying enzymes[epoxide hydrolase and glutathione-S-transferase(GST)] and intracellular organelles (microsomes and cytosol). In addition we also tested the suppression of DNA adducts formation by GSH and NAC. The results are summerized as follow. 1. The microsomes and cytosol which contain epoxide hydrolase and GST, respectively, decreased the mutagenicity of VCO (74% and 95%, respecfivel), and NPO (35% and 93%, respectively). The nucleophilic GSH and NAC decreased the mutagenicity by 86% (VCO) and 80% (NPO), 76% (VCO) and 40% (NPO), respectively. 2. The purified epoxide hydrolase decreased the mutagenicity of two epoxides in a dose-dependent manner, and GSH also decreased the mutagenicity in the presence of GST. 3. Formation of two DNA adducts, 7-(2'-oxoethyi)guanine (OEG) and N2,3-ethenoguanine(EG), were compared in the presence of calf thymus DNA and epoxide (VCO or NPO) in vitro system. The amounts of DNA adducts were decreased in the presence of GSH (25% and 29% in VCO, 32% and 29% in NPO), and NAC (14% and 16% in VCO, 21% and 11% in NPO), respectively. From these results, it is concluded that the ultimate carcinogenic metabolites, VCO and NPO, can be made in the body, but much of them may be inactivated and detoxified by the nucleophilic GSH, NAC and detoxifying enzymes (epoxide hydrolase and GST). Therefore, by these mechanism, the formation of DNA adducts and mutagenic activities of these two epoxides may be lowered in vivo.

• Toxicological Research
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• v.13 no.3
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• pp.257-263
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• 1997

Pentachlorophenol(PCP) which ks widely used in wood preservation, pulp and paper mills, has led to a substantial envirortmental contamination. To get the reliable data for the effective health risk assessment with PCP, covalent binding potential of PCP to cellular macromolecules and glutathione(GSH) was investigated after intraperitoneal administration of $^{14}C-PCP$ to rats. PCP metabolites were able to bind covalently to serum albumin and hepatic protein in a dose- and time-dependent manner. Hepatic protein adducts of PCP metabolites were increased as a function of cytochrome P-450 activities, whereas, albumin adducts significantly decreased. Covalent binding of PCP metabolites with DNA or hemoglobin was not observed. GSH levels in liver tissue decreased over 12hrs, however, the level was recovered after 48hrs. Tetrachloro-1,4-benzoquinone (1,4-TCBQ), one of the most reactive PCP metabolites, conjugated with GSH very rapidly. Base on our results, we could conclude that PCP metabolized to reactive electrophilic metabolites by cytochrome P-450 isoenzymes and conjugated rapidly with neighboring protein or nonprotein sulfhydryl before reacting with DNA or hemoglobin. We propose that albumin adducts and mercapturic acids of PCP metabolites can be used good biomarker of recent PCP exposure.

• Proceedings of the PSK Conference
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• 2002.10a
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• pp.359.1-359.1
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• 2002

In mammalian species, CpG dinucleotides are highly methylated with 60-90% methylation at the 5-position of cytosine. The pattern of DNA methylation in a cell dramatically affects the function of the DNA by switching genes on or off. Abnormal methylation events occur during aging and in the development of many cancers. Methylated CpG was reported recently to affect the reactivity of agents (mitomycin C and benzo ［a］pyrenediolepoxide) that can fromguanine adducts in DNA. (omitted)

• Korean Journal of Food Science and Technology
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• v.24 no.4
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• pp.367-370
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• 1992

Aflatoxins are highly carcinogenic agents consistantly found as contaminants in human food supplies in many areas of the world and epidemiologically linked to incidence of human liver cancer. To examine the carcinogenic action of aflatoxin $B_1$, aflatoxin $B_1-DNA$ adducts were chemically synhtesized with the reaction of 20 mg calf thymus DNA, and 8 mg standard aflatoxin $B_1$. Since DNA molecule was too large for analysis, it was fragmented by acid hydrolysis and heat. The fragmented aflatoxin $B_1-DNA$ adducts were selectively concentrated by immunoaffinity column procedure and confirmed by HPLC method. The main component was aflatoxin $B_1-guanine$ adduct, which was quantatively measured as 5.2 mg aflatoxin $B_1$.

• Proceedings of the Korean Society of Toxicology Conference
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• 2001.10a
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• pp.24-25
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• 2001

Persistent cellular oxidative stress and enhanced lipid peroxidation (LPO) of PUFAs, leading to macromolecular damage and disruption of signaling pathways, are implicated in the development of human malignancies and other chronic degenerative diseases. LPO generates by oxidation of linoleic acid (LA) or arachidonic acid ($\omega$ -6 PUPAs) reactive aldehydes, such as trans-4-hydroxy-2-nonenal, which form etheno $\varepsilon$ -DNA adducts in a variety of human tissues and thus can contribute to diet-related cancers.(omitted)

• Proceedings of the Korea Environmental Mutagen Society Conference
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• 2004.05a
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• pp.49-53
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• 2004